Fluorescence-Based Detection of Natural Transformation in Drug-Resistant Acinetobacter baumannii

Journal of Bacteriology
Anne-Sophie GodeuxXavier Charpentier

Abstract

Acinetobacter baumannii is a nosocomial agent with a high propensity for developing resistance to antibiotics. This ability relies on horizontal gene transfer mechanisms occurring in the Acinetobacter genus, including natural transformation. To study natural transformation in bacteria, the most prevalent method uses selection for the acquisition of an antibiotic resistance marker in a target chromosomal locus by the recipient cell. Most clinical isolates of A. baumannii are resistant to multiple antibiotics, limiting the use of such selection-based methods. Here, we report the development of a phenotypic and selection-free method based on flow cytometry to detect transformation events in multidrug-resistant (MDR) clinical A. baumannii isolates. To this end, we engineered a translational fusion between the abundant and conserved A. baumannii nucleoprotein (HU) and the superfolder green fluorescent protein (sfGFP). The new method was benchmarked against the conventional antibiotic selection-based method. Using this new method, we investigated several parameters affecting transformation efficiencies and identified conditions of transformability one hundred times higher than those previously reported. Using optimized transformation...Continue Reading

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Citations

Aug 29, 2019·Proceedings of the National Academy of Sciences of the United States of America·Isabelle DurieuxXavier Charpentier
Mar 6, 2021·Current Microbiology·Ulrike BlaschkeGottfried Wilharm
Jul 8, 2021·Applied and Environmental Microbiology·Félix BornierXavier Charpentier

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Datasets Mentioned

BETA
AB5075

Methods Mentioned

BETA
flow cytometry
fluorescence microscopy
PCR
protein folding
genetic modification
PCRs

Software Mentioned

BLAST

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